Characterization of Tilapia (Oreochromis niloticus) aldehyde reductase (AKR1A1) gene, promoter and expression pattern in benzo-a-pyrene exposed fish

IF 2.8 4区医学 Q2 TOXICOLOGY

Toxicology Mechanisms and Methods Pub Date : 2017-01-02 DOI:10.1080/15376516.2016.1238529

A. Hassanin, Y. Kaminishi, T. Itakura

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引用次数: 3

Abstract

Abstract This study planned to isolation and characterization of AKR1A1 cDNA from Bap injected nile tilapia (Oreochromis niloticus), comparison of its characteristic structures with those of other species, characterization of AKR1A1 gene and promoter, and investigation of AKR1A1 mRNA expression in various organs of Bap injected tilapia. The cDNA was 1172 bp long which includes an open reading frame of 975 bp encoding a 324 amino acids protein and a stop codon. The sequence showed 3' and 5' non-coding regions of 179 and 18 bp. The amino acid sequence of O. niloticus AKR1A1 shows similarities of 60, 60, 60.6, 61.2 62.2, and 57.8% with mouse AKR1A1, Norway rat AKR1A1, zebrafish AKR1A1, African clawed frog AKR1A1, human, and yellow perch AKR1A1, respectively. Nucleotide sequence investigation of AKR1A1 gene and 5′-flanking region showed that the structural gene and the 5′-flanking region were approximately 2975 bp and 4006 bp in length, respectively. The protein-coding region contained eight exons, and one additional upstream exon. Real-time polymerase chain reaction (PCR) results showed that the highest level of AKR1A1 expression was found in bile (108.7), followed by kidney (77.9), muscles (37.3), and liver (24.7). mRNA levels of AKR1A1 were almost negligible in gills (0.6) while no detectable (ND) constitutive expression was detected in gut. In conclusion, our results concluded that tilapia AKR1A1 is inducible by BaP and have a significant function in the metabolism of xenobiotics and, therefore, may used as biomarker in fish

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罗非鱼（Oreochromis niloticus）醛还原酶（AKR1A1）基因、启动子及其在苯并芘暴露鱼类中的表达特征

摘要本研究拟从Bap注射罗非鱼(Oreochromis niloticus)中分离和鉴定AKR1A1 cDNA，与其他物种的特征结构进行比较，鉴定AKR1A1基因和启动子，并研究AKR1A1 mRNA在Bap注射罗非鱼各器官中的表达情况。cDNA全长1172 bp，包含975 bp的开放阅读框，编码324个氨基酸的蛋白和一个停止密码子。序列显示3′和5′非编码区，分别为179和18 bp。niloticus AKR1A1氨基酸序列与小鼠AKR1A1、挪威大鼠AKR1A1、斑马鱼AKR1A1、非洲爪蛙AKR1A1、人类AKR1A1和黄鲈AKR1A1的相似性分别为60、60、60.6、61.2、62.2和57.8%。对AKR1A1基因和5 ' -flanking区域的核苷酸序列调查显示，结构基因和5 ' -flanking区域的长度分别约为2975 bp和4006 bp。蛋白质编码区包含八个外显子和一个额外的上游外显子。实时聚合酶链反应(Real-time polymerase chain reaction, PCR)结果显示，AKR1A1在胆汁中的表达量最高(108.7)，其次是肾脏(77.9)、肌肉(37.3)和肝脏(24.7)。在鳃中，AKR1A1的mRNA水平几乎可以忽略不计(0.6)，而在肠道中，没有检测到(ND)组成表达。综上所述，我们的研究结果表明，罗非鱼AKR1A1可被BaP诱导，在外源代谢中具有重要的功能，因此可以作为鱼类的生物标志物

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来源期刊

Toxicology Mechanisms and Methods TOXICOLOGY-

自引率

3.10%

发文量

期刊介绍： Toxicology Mechanisms and Methods is a peer-reviewed journal whose aim is twofold. Firstly, the journal contains original research on subjects dealing with the mechanisms by which foreign chemicals cause toxic tissue injury. Chemical substances of interest include industrial compounds, environmental pollutants, hazardous wastes, drugs, pesticides, and chemical warfare agents. The scope of the journal spans from molecular and cellular mechanisms of action to the consideration of mechanistic evidence in establishing regulatory policy. Secondly, the journal addresses aspects of the development, validation, and application of new and existing laboratory methods, techniques, and equipment. A variety of research methods are discussed, including: In vivo studies with standard and alternative species In vitro studies and alternative methodologies Molecular, biochemical, and cellular techniques Pharmacokinetics and pharmacodynamics Mathematical modeling and computer programs Forensic analyses Risk assessment Data collection and analysis.